Potential Applications of Forensic Microbiology. A Narrative Review.
DOI:
https://doi.org/10.5377/rcfh.v11i2.22421Keywords:
Forensic microbiology, Tanatomicrobiome, Necrobiome, Post-mortem Interval, Legal medicine, Bioterrorism, Human microbiomeAbstract
Objective
To conduct a narrative literature review to present the general aspects of the discipline and the most relevant potential applications of microbiology in the forensic field, emphasizing those related to the investigation of Postmortem Interval calculation, the investigation of causes of death, and bioterrorism.
Introduction
Until a few years ago, microbiology was not widely recognized as a forensic discipline; its most frequent applications were related to the identification of etiological agents of infections, in bioterrorism investigations, or in the investigation of sexual offenses, among others. Advances in analytical methodologies such as next-generation sequencing, genomics, metagenomics, bioinformatics, and artificial intelligence, as well as the impetus provided by the Human Microbiome Project, catapulted the potential applications of this discipline in the forensic field, ranging from human identification to geolocation applications, among others. However, despite the advances and potential applications, its practical applications and implementation in real-world cases are not always so dynamic, as some challenges arise for its routine use.
Methodology
A literature review was conducted in the PubMed, Google Scholar Labs, and ScienceDirect databases, covering the period from 1980 to 2026, using a combination of English and Spanish keywords: Forensic Microbiology, Thanatomicrobiome, Necrobiome, Postmortem Interval, Legal Medicine, Bioterrorism, Human Microbiome.
Results
Fifty-seven articles with full access and four relevant websites were selected.
Conclusion
Forensic microbiology is a powerful tool that requires further research, standardization, and validation of procedures for its routine adoption in forensic settings, especially in applications such as calculating the Postmortem interval and investigating causes of death.
Downloads
56
HTML (Español (España)) 3
References
1.- Budowle B, Schutzer SE, Einseln A, Kelley LC, Walsh AC, Smith JA, et al. Public health. Building microbial forensics as a response to bioterrorism. Science. 2003;301(5641):1852-3. doi: 10.1126/science.1090083.
2.- Franceschetti L, Lodetti G, Blandino A, Amadasi A, Bugelli V. Exploring the role of the human microbiome in forensic identification: opportunities and challenges. Int J Legal Med. 2024 ;138(5):1891-1905. Disponible en: https://doi.org/10.1007/s00414-024-03217-z
3.- Tozzo P, D'Angiolella G, Brun P, Castagliuolo I, Gino S, Caenazzo L. Skin microbiome analysis for forensic human identification: what do we know so far? Microorganisms. 2020 ;8(6):873. Disponible en: https://doi.org/10.3390/microorganisms8060873.
4.- Díez López C, Vidaki A, Kayser M. Integrating the human microbiome in the forensic toolkit: Current bottlenecks and future solutions. Forensic Sci Int Genet. 2022;56:102627. DOI:10.1016/j.fsigen.2021.102627
5.- Budowle B, Harmon R. HIV legal precedent useful for microbial forensics. Croat Med J. 2005 ;46(4):514-21. Disponible en: https://pubmed.ncbi.nlm.nih.gov/16100753/
6.- Zhu Y, Villalba JA, Kasten J, DeLeon Carnes M, Bhatnagar J, Zumwalt R. Sudden death with lyme disease myocarditis: a case report. Am J Forensic Med Pathol. 2025;46(4):319-322.
7.- Stassi C, Mondello C, Baldino G, Ventura Spagnolo E. Post-mortem investigations for the diagnosis of sepsis: a review of literature. Diagnostics (Basel). 2020;10(10):849. doi: 10.3390/diagnostics10100849.
8.- Fernández-Rodríguez A, Alberola J, Cohen MC. Análisis microbiológico post mórtem. Enferm Infecc Microbiol Clin . 2013 ;31(10):685-91. Disponible en: https://ln.run/dPawj
9.- Wang X, Le C, Jin X, Feng Y, Chen L, Huang X, et al. Estimating postmortem interval based on oral microbial community succession in rat cadavers. Heliyon. 2024;10(11):e31897. doi: 10.1016/j.heliyon.2024.e31897.
10.- Haarkötter C, Saiz M, Gálvez X, Medina-Lozano MI, Álvarez JC, Lorente JA. Usefulness of microbiome for forensic geolocation: a review. Life (Basel) . 2021 ;30;11(12):1322. Disponible en: https://pmc.ncbi.nlm.nih.gov/articles/PMC8707258/
11.- Cho HW, Eom YB. Forensic analysis of human microbiome in skin and body fluids based on geographic location. Front Cell Infect Microbiol. 11:695191. doi: 10.3389/fcimb.2021.695191.
12.- Dobay A, Haas C, Fucile G, Downey N, Morrison HG, Kratzer A, et al. Microbiome-based body fluid identification of samples exposed to indoor conditions. Forensic Sci Int Genet. 40:105–113. doi: 10.1016/j.fsigen.2019.02.010
13.- Huang L, Xu C, Yang W, Yu R. A machine learning framework to determine geolocations from metagenomic profiling. Biol Direct. 2020;15(1):27. doi: 10.1186/s13062-020-00278-z
14.- Clarke T, Brinkac L, Greco C, Alleyne AT, Carrasco P, Inostroza C, et al. Sampling from four geographically; divergent young female populations demonstrates forensic geolocation potential in microbiomes. Sci Rep. 2022. 12(1):18547. Disponible en: https://pubmed.ncbi.nlm.nih.gov/36329122/
15.- Lei Y, Li M, Zhang H, Deng Y, Dong X, Chen P, et al. Comparative analysis of the human microbiome from four different regions of China and machine learning-based geographical inference. mSphere. 2025;10(1): e0067224. doi: 10.1128/msphere.00672-24.
16.- Fernández-Huerta M, Zarzuela F, Barberá MJ, Arando M, Esperalba J, Rodríguez V, et al. Sexual transmission of intestinal parasites and other enteric pathogens among men who have sex with men presenting gastrointestinal symptoms in an sti unit in barcelona, spain: a cross-sectional study. Am J Trop Med Hyg. 2019;101(6):1388-91. DOI: 10.4269/ajtmh.19-0312.
17.- Costello EK, Lauber CL, Hamady M, Fierer N, Gordon JI, Knight R. Bacterial community variation in human body habitats across space and time. Science. 2009;326(5960):1694-7. DOI: 10.1126/science.1177486
18.-Suvitha MR, Afrin Sona AS, Jose L, Priyatha CV, Siva Prasad MS. Application of microbiome in forensics: a critical review. J Pure Appl Microbiol. 2025;19(4):2441-56. doi: 10.22207/JPAM.19.4.37
19.- Dixon R, Egan S, Payne M, Mullally C, Chapman B. La transferencia bacteriana durante las relaciones sexuales como herramienta de detección forense. Iscience. 2025;28(2):111861
20.- Fenner A. Sexual transfer of bacteria for forensic use. Nat Rev Urol. 2025 ; 22(5):253. Disponible en: https://doi.org/10.1038/s41585-025-01037-3
21.- Tambuzzi S, Maciocco F, Gentile G, Boracchi M, Bailo P, Marchesi M, et al. Applications of microbiology to different forensic scenarios - a narrative review. J Forensic Leg Med. 2023;98:102560
22.- Benítez-Pérez MO, Reyes-Roque AC, Gómez-Pacheco R, Calderón-Medina NA, Moreira-Silverio B, Godoy-León R. Armas biológicas: antecedentes históricos, producción y usos en diferentes etapas de la guerra biológica. Arch méd Camagüey. 2024 [Consultado 15 de diciembre de 2025]; 28: e10132. Disponible en: https://revistaamc.sld.cu/index.php/amc/article/view/10132
23.-Bosch X. (1998). Hepatitis C outbreak astounds Spain. 351(9113), 1–. doi:10.1016/s0140-6736(05)79465-4
24.- Ravel J, Jiang L, Stanley ST, Wilson MR, Decker RS, Read TD, et al. The complete genome sequence of Bacillus anthracis Ames "Ancestor". J Bacteriol. 2009;191(1):445-6. DOI: 10.1128/JB.01347-08
25.- Budowle B, Murch R, Chakraborty R. Microbial forensics: the next forensic challenge. Int J Legal Med. 2005;119(6):317-30. doi: 10.1007/s00414-005-0535-y.
26.- NIH HMP Working Group; Peterson J, Garges S, Giovanni M, McInnes P, Wang L, et al. The NIH human microbiome project. Genome Res. 2009;19(12):2317-23. doi: 10.1101/gr.096651.109.
27.- National Institutes of Health. Human Microbiome Project (HMP). Washington: NIH; 2025. Disponible en: https://commonfund.nih.gov/hmp
28.- Berg G, Rybakova D, Fischer D, Cernava T, Vergès MC, Charles T, et al. Microbiome definition re-visited: old concepts and new challenges. Microbiome. 2020;8(1):103. DOI: 10.1186/s40168-020-00875-0
29.- Baylor College of Medicine. The human microbiome: the basics. Houston: Baylor College of Medicine; sf. Disponible en: https://www.bcm.edu/departments/molecular-virology-and-microbiology/emerging-infections-and-biodefense/microbiome
30.- The Integrative HMP, Research Network Consortium. The integrative human microbiome project. Nature. 2019; 569(7758):641-48. Disponible en: https://pmc.ncbi.nlm.nih.gov/articles/PMC6784865/
31.- Heidrich V, Valles-Colomer M, Segata N. Human microbiome acquisition and transmission. Nat Rev Microbiol. 2025 ;23(9):568-84. Disponible en: https://doi.org/10.1038/s41579-025-01166-x
32.- García MG, Pérez-Cárceles MD, Osuna E, Legaz I. Impact of the human microbiome in forensic sciences: a systematic review. Appl Environ Microbiol. 2020 ;86(22):e01451-20. Disponible en: https://doi.org/10.1128/AEM.01451-20
33.- Dass M, Abbai NS, Ghai M. The human skin microbiome: factors affecting individuality and application in forensic investigations. Int J Legal Med. 2026 ;140(2):1067-84. Disponible en: https://doi.org/10.1007/s00414-025-03610-2.
34.- Metcalf JL, Wegener Parfrey L, González A, Lauber CL, Knights D, Ackermann G, et al. A microbial clock provides an accurate estimate of the postmortem interval in a mouse model system. Elife. 2013; 15;2:e01104. Disponible en: https://pmc.ncbi.nlm.nih.gov/articles/PMC3796315/
35.- Teixeira MJ, Barbosa DJ, Dinis-Oliveira RJ, Freitas AR. Redefining postmortem interval estimation: the need for evidence-based research to bridge science and justice. Front Microbiol. 2025:16:1646907. doi: 10.3389/fmicb.2025.1646907
36.- Y K, Isukapatla AR. Postmortem microbiome dynamics: review of forensic microbial clock. J Forensic Leg Med . 2026 ;117:103024. Disponible en: https://doi.org/10.1016/j.jflm.2025.103024
37.- Oliveira M; Amorim A. Microbial forensics: new breakthroughs and future prospects. Appl Microbiol Biotechnol. 2018;102(24):10377-91. doi:10.1007/s00253-018-9414-6
38.- Wu Z, Guo Y, Hayakawa M, Yang W, Lu Y, Ma J, et al. Artificial intelligence-driven microbiome data analysis for estimation of postmortem interval and crime location. Front Microbiol. 2024;15:1334703. doi: 10.3389/fmicb.2024.1334703.
39.- Can I, Javan GT, Pozhitkov AE, Noble PA. Distinctive thanatomicrobiome signatures found in the blood and internal organs of humans. J Microbiol Methods. 2014;106:1-7. doi:10.1016/j.mimet.2014.07.026
40.- Tuomisto S, Karhunen PJ, Vuento R, Aittoniemi J, Pessi T. Evaluation of postmortem bacterial migration using culturing and real-time quantitative PCR. J Forensic Sci. 2013;58(4):910-6. doi: 10.1111/1556-4029.12124.
41.- Lei Y, Li M, Zhang H, Deng Y, Dong X, Chen P, et al. Comparative analysis of the human microbiome from four different regions of China and machine learning-based geographical inference. mSphere. 2025;10(1):e0067224. doi: 10.1128/msphere.00672-24.
42.- Tsokos M, Püschel K. Postmortem bacteriology in forensic pathology: diagnostic value and interpretation. Leg Med (Tokyo). 2001;3(1):15-22. doi: 10.1016/s1344-6223(01)00002-5.
43.- Gunn A, Pitt SJ. Microbes as forensic indicators. Trop Biomed. 2012 Consultado];29(3):311-30. Disponible en: https://www.msptm.org/files/311_-_330_Alan_Gunn.pdf
44.- Nodari R, Arghittu M, Bailo P, Cattaneo C, Creti R, D'Aleo F, et al. Forensic microbiology: when, where and how. Microorganisms. 2024;12(5):988. doi: 10.3390/microorganisms12050988
45.- Ventura Spagnolo E, Stassi C, Mondello C, Zerbo S, Milone L, Argo A. Forensic microbiology applications: a systematic review. Leg Med (Tokyo). 2019;36:73-80. doi: 10.1016/j.legalmed.2018.11.002.
46.- Speruda M, Piecuch A, Borzęcka J, Kadej M, Ogórek R. Microbial traces and their role in forensic science. J Appl Microbiol . 2022 ;132(4):2547-2557. Disponible en: https://pubmed.ncbi.nlm.nih.gov/34954826/
47.- Uchiyama T, Kakizaki E, Kozawa S, Nishida S, Imamura N, Yukawa N. A new molecular approach to help conclude drowning as a cause of death: simultaneous detection of eight bacterioplankton species using real-time PCR assays with TaqMan probes. Forensic Sci Int. 2012;222(1-3):11-26. DOI:10.1016/j.forsciint.2012.04.029
48.- Camatti J, Bonasoni MP, Santunione AL, Cecchi R, Radheshi E, Carretto E. Postmortem microbiology in forensic diagnostics: interpretation of infectious causes of death and emerging applications. Diagnostics. 2026 ;19;16(2):325. Disponible en: https://doi.org/10.3390/diagnostics16020325
49.- Fernández-Rodríguez A, Cohen MC, Lucena J, Van de Voorde W, Angelini A, Ziyade N, et al. How to optimise the yield of forensic and clinical post-mortem microbiology with an adequate sampling: a proposal for standardisation. Eur J Clin Microbiol Infect Dis. 2015;34(5):1045-57.
50.- Oliveira M, Mason-Buck G, Ballard D, Branicki W, Amorim A. Biowarfare, bioterrorism and biocrime: A historical overview on microbial harmful applications. Forensic Sci Int . 2020 ;314:110366. Disponible en: https://pmc.ncbi.nlm.nih.gov/articles/PMC7305902/
51.- Oficina de Asuntos de Desarme de las Naciones Unidas. Armas biológicas . New York: ONU; sf . Disponible en: https://disarmament.unoda.org/es/our-work/weapons-mass-destruction/biological-weapons
52.- Marquina Díaz D, Sánchez JV, Santos de la Sen A. ¿Qué son las armas biológicas? un recorrido por su utilización a lo largo de la historia bélica . Madrid: Universidad Complutense de Madrid; sf. Disponible en: https://www.ucm.es/otri/noticias-que-son-las-armas-biologicas-un-recorrido-por-su-utilizacion-a-lo-largo-de-la-historia-belica
53.- López-Muñoz F. Shirō Ishii: el mayor criminal de guerra médico de la historia que nunca fue juzgado. BBC News Mundo . 21 marzo 2021 . Disponible en: https://www.bbc.com/mundo/noticias-internacional-56410365
54.- Mir TUG, Wani AK, Akhtar N, Sena S, Singh J. Microbial forensics: A potential tool for investigation and response to bioterrorism. Health Sci Rev . 2022 ;5:100068. Disponible en: https://doi.org/10.1016/j.hsr.2022.100068
55.- Murch RS. Bioattribution needs a coherent international approach to improve global biosecurity. Front Bioeng Biotechnol. 2015;3:80. doi: 10.3389/fbioe.2015.00080.
56.- Tackmann J, Arora N, Schmidt TSB, Rodrigues JFM, von Mering C. Ecologically informed microbial biomarkers and accurate classification of mixed and unmixed samples in an extensive cross-study of human body sites. Microbiome. 2018;6(1):192. doi: 10.1186/s40168-018-0565-6.
57.- Gouello A, Dunyach-Remy C, Siatka C, Lavigne JP. Analysis of microbial communities: an emerging tool in forensic sciences. Diagnostics (Basel) . 2021 ;12(1):1. Disponible en: https://doi.org/10.3390/diagnostics12010001
58.- Sung J, Rajendraprasad SS, Philbrick KL, Bauer BA, Gajic O, Shah A. The human gut microbiome in critical illness: disruptions, consequences, and therapeutic frontiers. J Crit Care. 2024;79:154436. DOI: https://doi.org/10.1016/j.jcrc.2023.154436.
59.- Singh H, Clarke T, Brinkac L, Greco C, Nelson KE. Forensic microbiome database: a tool for forensic geolocation meta-analysis using publicly available 16s rRNA microbiome sequencing. Front Microbiol. 2021;12:644861. DOI:.10.3389/fmicb.2021.644861
60.- Logan AC, Cordell B, Pillai SD, Robinson JM, Prescott SL. From Bacillus Criminalis to the Legalome: Will Neuromicrobiology Impact 21st Century Criminal Justice?. Brain Sci . 2025 ;15(9):984. Disponible en: https://doi.org/10.3390/brainsci15090984
61.- Zhang J, Yu D, Zhang L, Wang T, Yan J. Environmental microbiota from substrate may interfere with microbiome-based identification of forensically relevant body fluids: a pilot study. Forensic Sci Int Genet . 2025 ;74:103170. Disponible en: https://doi.org/10.1016/j.fsigen.2024.10314
Published
How to Cite
Issue
Section
License
Copyright (c) 2026 Mireya Matamoros Zelaya, Carolina Inostroza
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
El autor conserva los derechos de autor bajo los terminos de una licencia CC NC 4.0
